在这项研究中,进行了两个实验来评估褪黑素对人体外胚胎质量的影响和作用。
实验一:共纳入42例重复劣质胚胎患者,共181个卵母细胞回收周期。IVF后,对于同一个病人,将MT周期组(向培养基中添加10-7M褪黑激素;n=48)与先前的非MT周期组(n=133)进行比较,其次是体外培养到囊胚期和胚胎移植。31例患者进行65例胚胎移植,包括24个MT胚胎移植,41非MT胚胎移植。比较两组的周期结局。实验II:在IVF后第3天玻璃化的总共143个多余的人类卵裂期胚胎(来自非重复质量差的胚胎患者)被加热并随机分为两组:褪黑激素组(10-7M褪黑激素添加到培养基中;n=71)和对照组(n=72),然后培养72小时。囊胚率和高质量囊胚率,培养基的活性氧(ROS)水平以及胚胎GPX1,CAT,Mn-SOD,Cu/Zn-SOD,分析BCL-2、BAX基因表达水平。
实验一:结果表明,MT周期组的第3天优质胚胎率(29.6%vs.19.5%)明显高于非MT周期组(P<0.05)。褪黑素治疗组的可用囊胚率(17.1%vs.12.7%)和临床妊娠率(25.0%vs.17.1%)有升高趋势(P>0.05)。实验二:结果表明,褪黑素给药组囊胚率明显高于对照组(42.25%vs.26.38%,P<0.05)。高质量囊胚率无显著差异。此外,定量PCR显示褪黑素处理后CAT的表达明显上调(P<0.05),而GPX1、Mn-SOD的表达无显著差异,Cu/Zn-SOD,BAX和BCL-2基因以及ROS的水平。
这些数据表明,在培养基中补充褪黑激素将提高重复劣质胚胎患者的第3天优质胚胎率,并提高玻璃化温热卵裂期胚胎的囊胚率,提示褪黑素干预可能为IVF失败提供潜在的抢救策略。
标识符[ChiCTR2200059773]。
In this
study, two experiments were performed to assess the effect and the role of melatonin on human in vitro embryo quality.
Experiment I: A total of 42 repeated-poor-quality-embryo patients were enrolled, with a total of 181 oocytes retrieval cycles. After IVF, for the same patient, the MT cycles group (10-7 M melatonin added to the culture medium; n=48) were compared with the previous non-MT cycles group (n=133), following by in vitro culture to blastocyst stage and embryo transfer. 31 patients were transplanted with 65 embryo transfer, including 24 MT embryo transfer, 41 non-MT embryo transfer. Cycle outcomes were compared between the two groups. Experiment II:A total of 143 supernumerary human cleavage-stage embryos (from non-repeated-poor-quality-embryo patients) vitrified on Day 3 after IVF were warmed and randomized into two groups: melatonin group (10-7 M melatonin added to the culture medium; n=71) and control group (n=72), and then cultured for 72 h. Rate of blastocyst and high-quality blastocyst, reactive oxygen species (ROS) levels of culture media as well as embryonic GPX1, CAT, Mn-SOD, Cu/Zn-SOD, BCL-2, BAX gene expression levels were analyzed.
Experiment I: Results showed that the rate of Day 3 high-quality embryos (29.6% vs.19.5%) in the MT cycles group was significantly higher than that in the non-MT cycles group (P<0.05). The rate of available blastocysts (17.1% vs.12.7%) and clinical pregnancy rate (25.0% vs.17.1%) were in tendency higher in the group treated with melatonin (P>0.05). Experiment II:Results showed that the blastocyst rates in the melatonin administered group were significantly higher than in control group (42.25% vs.26.38%, P<0.05). There were no significant differences in high-quality blastocyst rates. In addition, quantitative PCR showed that the expression of CAT was significantly upregulated by melatonin treatment (P<0.05), while there were no significant differences in the expression of GPX1, Mn-SOD, Cu/Zn-SOD, BAX and BCL-2 gene as well as the levels of ROS.
These data showed that melatonin supplement in the culture medium will improve Day 3 high-quality embryos rate of repeated-poor-quality-embryo patients and improve blastocyst rate of vitrified-warmed cleavage-stage embryos, suggesting that melatonin intervention may provide a potential rescue strategy for IVF failures.
identifier [ChiCTR2200059773].